The Toxic Effect of Zinc Oxide Nanoparticles on the Terrestrial Slug Lehmannia Nyctelia (Gastropoda-Limacidae) Wafaa A

Mohammad et al. The Journal of Basic and Applied Zoology (2021) 82:34 The Journal of Basic https://doi.org/10.1186/s41936-021-00214-1 and Applied Zoology RESEARCH Open Access The toxic effect of zinc oxide nanoparticles on the terrestrial slug Lehmannia nyctelia (Gastropoda-Limacidae) Wafaa A. Mohammad1, Safaa M. Ali2* , Nasser Farhan3 and Shimaa Mohamed Said2 Abstract Background: Chemicals have deleterious effect on the environment. The wide use of nanomaterials as products for plant protection, fertilizers, and also in water purification leads to the release of these materials to the environment. Terrestrial gastropods including snails and slugs have the ability to accumulate heavy metals in their bodies. The present study evaluates the toxic effect of zinc oxide nanoparticles on the terrestrial slug Lehmannia nyctelia. Zinc oxide nanoparticles (ZnO NPs) were prepared by thermal decomposition method. ZnO NPs are characterized by X- ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and ultraviolet-visible spectroscopy (UV). Slugs were treated with three different concentrations of ZnO NPs. Results: A total of three animals died by the end of the experiment. Many histological alterations were detected after exposure to different concentration of ZnO NPs. Conclusions: The obtained histological alterations prove the toxic effect of ZnO NPs on the animal under study. Background effects such as toxicity of non-target organisms, contam- Molluscs are considered as one of the important animals ination of food and environment (Jeong et al., 2012). in the food chain and noticed as agricultural pests. Toxic Terrestrial gastropods are one of the most efficient ac- chemicals produce harmful effect on terrestrial gastro- cumulators of metals and they are considered as bio- pods at the cellular level (Boer et al., 1995; Hernadi indicators to environmental pollution (Pihan & de Vau- et al., 1992). At the same time, they possess special fleury, 2000; Snyman et al., 2005; Viard et al., 2004). Ter- mechanisms to get rid of several harmful chemicals restrial snails and slugs have a great capacity of the which accumulated in their tissues (Ferner, 2001). accumulation and magnification of heavy metals and are Slugs are considered as a great threat to many of the considered important species to monitor the bioavail- temperate horticultural crops. Seedlings are very sensi- ability of metallic components in the soil if they com- tive to slug as if they cause a little amounts of damage to pared to other invertebrates (Coeurdassier et al., 2000; the growing tips of seedlings, this will lead to plant death Lanno & Mc Carty, 1997; Wensen et al., 1994). (Moens & Glen, 2002; Nash et al., 2007). Recently, the industrial development in agricultural Slug Lehmannia valentiana is a pest which result in field increased rapidly and leads to inorganic and organic many troubles to greenhouse. The common method to contamination from harmful heavy metals and chemicals control this slug is the chemical molluscicides. The to the terrestrial and aquatic ecosystems. These chemi- chemical molluscicides may cause undesirable side cals show deleterious effect on aquatic and terrestrial environment (Davidson et al., 2011; Pack et al., 2014; Sanchez, 2008). * Correspondence: [email protected] Nanomaterials were widely used in agriculture during 2Zoology Department, Faculty of Science, Assiut University, Assiut, Egypt the last 10 years (Buzea & Pacheco, 2017; Gogos et al., Full list of author information is available at the end of the article © The Author(s). 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. Mohammad et al. The Journal of Basic and Applied Zoology (2021) 82:34 Page 2 of 9 2012). They were using as plant protection products, fer- applying Scherrer’s equation: D =Kλ/βcosθ, where K is tilizers, water purification, for soil improvement, and the shape factor whose value is taken as 0.89, λ is the pollutant remediation (Parisi et al., 2015). In many cases, wavelength of Cu Kα radiation (λ = 1.54178), and β is nanoparticles (NPs) are applied to the soil directly as a the corrected full width at half maximum (FWHM) of fertilizer or in the form of an aerosol (Sturikova et al., the diffraction peak and θ is the diffracting angle. 2018) while seeds are mostly soaked in aqueous NP sus- pension (Lin & Xing, 2007; Segatto et al., 2018). In the Fourier transform infrared spectroscopy (FTIR): future, technological application of NPs may be un- Infrared spectra were recorded on a Thermo Fisher– desired as they are released to the environment and dif- model: Nicolet iS10 FTIR spectrometer in a wave num- − fused in the soil due to their small size (Kaegi et al., ber range 4000–500 cm 1. 2008). Zinc oxide nanoparticles (ZnO nanoparticles) are in- SEM and TEM teresting materials due to their unique electrical and op- The morphology of the ZnO NPs powder sample was in- tical properties, their wide-band semiconductor, and vestigated by scanning electron microscopy (SEM; JEOL they are suitable for many applications (Mishra et al., [model: JSM 5400LV]) and transmission electron mi- 2011). ZnO NPs are useful in environmental remediation croscopy (TEM; JEOL [model: JEM-100 CXII]). due to their high capabilities for degradation and elimin- ation of pollutants in air or water due to their strong UV-visible spectroscopy physical adsorption and their chemical catalytic proper- The UV-vis absorption measurements of ZnO NPs were ties (Jing et al., 2001). Zn ONPs are used as antimicro- recorded in the range 200–800 nm by PerkinElmer; bial agent, and also used in cosmetics, sunscreens, and model: LAMBDA 750 UV/Vis/NIR Spectrophotometer. coatings (Ali et al., 2012). Due to the increasing use in The synthesized ZnO NPs powder sample was dispersed consumer products, ZnO NPs will be found in the at- in ethanol and optical characterizations were executed. mospheric, terrestrial, and aquatic environments (Ali et al., 2012; Sales, 2013; Service, R.F., 2008). Collection of specimens The present study aimed to evaluate the toxic effect of The experimental work was carried out using specimens ZnO NPs on theterrestrial slug Lehmannia nyctelia. of Lehmannia nyctelia, collected during autumn from Assiut University farm, Assiut Governorate, Egypt. Slugs Methods were kept in normal laboratory conditions, maintained Chemicals and reagents in plastic containers containing soil obtained from their All chemical and reagents purchased were analytical natural habitat and fed with fresh lettuce daily. grade. Zinc nitrate hydrate was purchased from Alfa Aesar (Ward Hill, MA), Ethanol, abs. 100% a.r. was pro- Experimental design and ZnO NPs exposure cured from Chem-Lab., Belgium. For the experiment, the acclimatized healthy slugs with an equal body weight (0.6 gm) were randomly distrib- Preparation of ZnO nanoparticles uted into four groups (control and three treated groups). The zinc oxide nanoparticles (ZnO NPs) were prepared Each group consists of 10 individuals and was kept in by thermal decomposition method as described by Shan- plastic containers containing the same weight of soil and kar et al. (2013). Three grams of zinc nitrate were taken each container covered with perforated cloth for ventila- and fired in a furnace (air atmosphere) for 3 h at 500 °C. tion. The control slug’s group fed on fresh lettuce It was further grinded in a mortar to make a fine powder impressed in 10 ml distilled water. of ZnO NPs. However, the treated groups were fed on fresh lettuce impressed in different concentrations of ZnO NPs (0.01 Characterization of ZnO NPs g/10 ml dw, 0.02/10 ml dw, 0.05/10 ml dw, modified ZnO NPs were characterized by X-ray diffraction (XRD), from Fahmy and Abdel-Ghaffar (2014). Exposure period Fourier transform infrared spectroscopy (FTIR), scan- was 10 days, during which the slugs were subjected twice ning electron microscopy (SEM), and ultraviolet-visible to the contaminated food with the previously mentioned spectroscopy (UV). concentrations of ZnO NPs. At the end of the experimental period, surviving slugs were used for histological X-ray diffraction (XRD) studies. Died animals were counted and removed. The structure of the nanoparticles was investigated by a Slugs were fixed in neutral formalin and prepared for Philips X-ray diffractometer (model PW 1720) at room histological study. Seven micrometers transverse sections temperature, with a step of 0.06°, in the range of 4° ≤ 2θ were prepared and stained with hematoxilin and eosin ≤ 89.89°. The average crystallite sizes were calculated by (HE). Sections were examined by light microscope. The Mohammad et al. The Journal of Basic and Applied Zoology (2021) 82:34 Page 3 of 9 organs under study are the digestive gland, salivary while Figure 2b presents a typical TEM image of ZnO glands, and hermaphroditic glands. NPs. It is clear that in this case a ZnO NPs was success- fully prepared. Results Characterization of ZnO nanoparticle The UV-Vis spectrum X-ray diffraction (XRD) The UV-Vis spectrum of ZnO NPs is shown in Fig. 2c. The ZnO NPs were characterized by X-ray diffraction Confirmation of the synthesized ZnO product in nano- (XRD).

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